RU2443425C1 - Method for making ozero lechebnoye mud extract for enhancing antiinflammatory and wound healing action - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to pharmaceutical industry, namely to a method for making a mud extract possessing wound healing action. The method for making the mud extract possessing wound healing action wherein a mud source is a native peloid feed - Ozero Lechebnoye mud which is extracted by three-stage filling with an aqueous solution of hydrochloric acid with vigorous stirring to produce a homogenous suspension that is followed by settling at room temperature, filtering micelles, driving off water in a rotary evaporator and sterilising under certain conditions.

EFFECT: method provides producing the mud with higher wound healing and anti-inflammatory actions.

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Description

The invention relates to medicine and is aimed at creating the basics of biologically active organic and inorganic substances for the subsequent development of peloid-containing drugs with anti-inflammatory and wound healing activity. As raw materials, medium-sulphide (FeS 0.15-0.3%) salt-saturated (mineralization> 250-400 g / dm ² ) silt muds from the Lechebnoe Lake of the Astrakhan Region are used.

A known method of obtaining a drug from sludge therapeutic mud, which can be taken as a prototype, which consists in the extraction of mud with water in the ratio

1: 2.5 in the presence of an isotonic concentration of sodium chloride, infusion at room temperature, sedimentation, sterilization and filtration (I.A. Muraviev. Drug Technology. M., Medicine, 1971, p.275). However, the known method does not provide therapeutic mud with a high concentration of biologically active substances and does not provide a pharmacologically active agent.

The aim of the invention is to optimize the method of obtaining extracts from mud "Lake" Healing "to increase specific anti-inflammatory and wound healing activity.

The technical result is the production of hydrochloric acid mud extract with anti-inflammatory and wound healing effects, exceeding the strength of the action of the aqueous extract obtained from similar raw materials.

The inventive method is characterized by the following set of operations: extraction of native mud with natural humidity 38% aqueous (1: 1) hydrochloric acid solution, threefold filling at a ratio of 3-4 volumes per 1 kg of raw material at 25-30 ° C with vigorous stirring to obtain a homogeneous suspension . Miscella is filtered after 12 hours of settling at room temperature. Water is distilled off on a rotary evaporator at t 50-60 ° C and a residual pressure of 10-15 mm Hg. The resulting extract is sterilized for 2-3 hours at 125-130 ° C. Of the above operations, distinctive from the prototype is the following: the use of an aqueous solution of hydrochloric acid, threefold pouring at a ratio of 3-4 volumes.

The set of operations of the claimed method allows to achieve an increase in the biological properties of the mud extract, as demonstrated in the examples for the study of anti-inflammatory and wound healing activity.

The anti-inflammatory effect of the native drug in the form of hydrochloric acid extract was studied in comparison with an aqueous mud extract in experiments on 40 Wistar rats. The anti-inflammatory effect of the extract was evaluated by its effect on the exudative phase of inflammation caused by subcutaneous injection of 0.1 ml of the phlogogenic agent formalin (2% p-p) in anesthetized animals with ether. After the introduction of formalin, the size of the inflammatory focus was noted every hour. The treatment with mud extract on the first day was carried out 2 hours, 3 hours and 4 hours after the induction of the inflammatory reaction, the next day in the morning (11.00) and evening hours (16.00). The inflammatory infiltrate was measured after 1 h, 2 h, 12 h, 24 h, 36 h, 48 h, 60 h and 72 h after the first application of the test drug. A bandage with a solution of mud extract was fixed to the back of the animal with a band-aid.

In the course of the study, it was found that the studied hydrochloric acid extract of the mud has a pronounced inhibitory effect on the exudative phase of the inflammatory process (table 1). After 1 h and 2 h after application, the area of induced edema decreased by 20 and 30%, respectively, while on the second day in 70% of the animals the signs of inflammation completely disappeared, in the remaining 30% of the animals, the area of inflammation decreased by more than 50%. On the 3rd day of observation, in 90% of the animals, the signs of the inflammatory reaction were completely reduced. On day 4, all animals were healthy, while in the control group and the comparison group (water mud extract) the inflammation zone decreased by no more than 40-50% compared with the first day of the experiment. Thus, the maximum (100%) antiphlogistic effect is manifested by the hydrochloric acid extract from the mud of Lake “Lechebnoe” by the end of 2 days of the development of the inflammatory reaction.

The wound healing effect of hydrochloric acid mud extract in comparison with its aqueous counterpart was studied in an experiment on 40 Wistar rats on a model of a chemical (intradermal injection of 0.3 ml of a 40% formalin solution) burn of surface tissues. Mud extract treatment was carried out by analogy with a series of experiments to study the anti-inflammatory effect of the extract. The effectiveness was judged by the time (in days) of the clinical healing of a burn defect (table 2).

24 hours after the introduction of the chemical inducer of superficial burns, a skin defect in the form of an inflammatory infiltrate with necrotic changes covered with fibrinous plaque. In the control group of animals that were not treated with mud extract, a slow decrease in the size of inflammatory infiltrate, defect zone and fibrinous plaque was observed; on day 10, 20% of the animals showed no signs of necrotic changes. In the experimental group of animals, on the second day of the beginning of the application of hydrochloric acid extract, the clinical picture sharply differed from the group of animals with a burn model. On day 4, the following was observed in 60% of animals: fibrinous plaque was insignificant, inflammatory infiltrate and necrotic tissue defect were absent. By 6 days, all animals showed no signs of burns. In the comparison group on day 6, 40% of the animals receiving the water extract applications did not show signs of necrosis; 100% wound healing effectiveness of the aqueous extract was manifested on the 9th day.

Table 1 Anti-inflammatory effect of extracts from mud “Lakes“ Healing ” Area of inflammatory infiltrate (mm) after formalin induction of an inflammatory reaction Number of individuals 1 hour 2 hours 3 hours 4 hours 12 hours 24 hours 36 hours 48 hours Inflammation model (0.1 ml of 2% formalin solution subcutaneously in the back) one 1.4 × 0.5 1.6 × 0.5 1.6 × 0.5 1.6 × 0.5 1.0 × 0.7 0.9 × 0.6 0.8 × 0.5 0.7 × 0.5 2 0.5 × 1.8 1.0 × 1.8 1.0 × 1.8 1.0 × 1.8 0.8 × 0.8 0.7 × 0.7 0.6 × 0.6 0.5 × 0.6 3 2.0 × 1.0 2.0 × 1.5 2.0 × 1.5 2.0 × 1.5 0.5 × 0.5 0.4 × 0.4 0.3 × 0.3 0.2 × 0.3 four 1.6 × 1.0 1.8 × 1.3 1.8 × 1.3 1.8 × 1.3 0.5 × 0.5 0.4 × 0.4 0.3 × 0.3 0.2 × 0.3 5 1.8 × 1.0 2.0 × 1.3 2.0 × 1.3 2.0 × 1.3 0.3 × 0.3 0.2 × 0.2 0.1 × 0.1 No signs of inflammation 6 1.5 × 1.5 2.0 × 1.5 2.0 × 1.5 2.0 × 1.5 0.2 × 0.2 0.1 × 0.2 0.1 × 0.1 7 2.0 × 1.0 2.0 × 1.3 2.0 × 1.3 2.0 × 1.3 0.6 × 0.3 0.5 × 0.2 0.4 × 0.1 0.3 × 0.1 8 1.7 × 1.0 2.0 × 1.2 2.0 × 1.2 2.0 × 1.2 0.8 × 0.5 0.7 × 0.4 0.6 × 0.3 0.5 × 0.2 Experience: model of inflammation (0.1 ml of 2% formalin solution subcutaneously in the back) + treatment with hydrochloric mud extract (according to the scheme indicated in the text) After each measurement, a mud extract is applied to the inflammation zone, previously rubbed one 1.8 × 1.2 1.8 × 1.5 1.5 × 1.3 1.3 × 1.1 No signs of inflammation No signs of inflammation No signs of inflammation No signs of inflammation 2 1.5 × 1.0 1.8 × 1.1 1.5 × 0.9 1.3 × 0.7 3 0.5 × 0.5 1.0 × 1.0 0.8 × 0.9 0.7 × 0.8 four 2.0 × 1.2 2.2 × 1.5 1.8 × 1.3 1.6 × 1.1 5 2.0 × 0.7 2.1 × 1.5 1.7 × 1.3 1.5 × 1.1 6 1.8 × 0.7 1.9 × 1.0 1.6 × 0.8 1.4 × 0.7 7 1.5 × 0.7 1.8 × 1.5 1.5 × 1.3 1.3 × 1.1 8 2.0 × 1.5 2.2 × 2.0 1.8 × 1.6 1.6 × 1.4 0.1 × 0.1 9 1.5 × 0.7 2.2 × 1.0 1.8 × 0.9 1.6 × 0.7 0.2 × 0.2 0.1 × 0.1 10 2.0 × 1.5 2.0 × 1.7 1.7 × 1.4 1.5 × 1.3 0.4 × 0.3 0.3 × 0.2 0.2 × 0.1 Experience: model of inflammation (0.1 ml of 2% formalin solution subcutaneously in the back) + treatment with an aqueous mud extract (according to the scheme indicated in the text) After each measurement, a mud extract is applied to the inflammation zone, previously rubbed one 2.0 × 1.5 2.0x1.6 1.6 × 0.5 1.6 × 0.5 0.6 × 0.5 0.5 × 0.4 0.3 × 0.4 0.3 × 0.2 2 2.0 × 1.5 2.2 × 1.7 1.0 × 1.8 1.0 × 1.8 0.5 × 0.5 0.4 × 0.4 0.3 × 0.4 0.3 × 0.2 3 1.0 × 1.0 1.7 × 1.0 2.0 × 1.5 2.0 × 1.5 0.5 × 0.7 0.4 × 0.5 0.4 × 0.5 0.4 × 0.3 four 2.1 × 2.0 2.3 × 2.2 1.8 × 1.3 1.8 × 1.3 0.5 × 0.4 0.4 × 0.4 0.3 × 0.4 0.3 × 0.3 5 2.0 × 1.2 2.0 × 1.6 2.0 × 1.3 2.0 × 1.3 0.2 × 0.4 0.2 × 0.4 No signs of inflammation No signs of inflammation 6 1.5 × 0.7 2.0 × 1.4 2.0 × 1.5 2.0 × 1.5 0.6 × 0.4 0.4 × 0.4 0.3 × 0.4 7 1.3 × 1.0 2.0 × 1.5 2.0 × 1.3 2.0 × 1.3 0.5 × 0.5 0.3 × 0.3 0.3 × 0.3 0.3 × 0.3 8 1.5 × 1.0 1.7 × 1.2 2.0 × 1.2 2.0 × 1.2 0.2 × 0.2 0.2 × 0.2 0.2 × 0.2 0.2 × 0.2 9 1.2 × 0.7 1.5 × 1.2 2.0 × 1.3 2.0 × 1.3 0.6 × 0.7 0.4 × 0.5 0.3 × 0.5 0.3 × 0.3 10 2.0 × 1.0 2.0 × 1.3 2.0 × 1.2 2.0 × 1.2 0.3 × 0.4 0.3 × 0.3 0.3 × 0.3 0.3 × 0.2

table 2 Wound healing effect of extracts from mud "Lakes" Healing " Area of necrotic defect (mm) after formalin induction of a chemical burn Number of individuals 1 day 2 days 3 days 4 days 5 days 6 day 7 days 8 day 9 days 10 days Model of a chemical burn (0.3 ml of a 40% solution of formalin intradermally in the back) one 1.2 × 1.2 1.5 × 1.2 1.3 × 1.2 1.2 × 1.2 1.0 × 1.0 1.0 × 0.8 0.9 × 0.7 0.8 × 0.6 0.7 × 0.5 0.6 × 0.5 2 1.1 × 1.6 1.3 × 1.6 1.1 × 1.6 1.1 × 1.6 0.9 × 1.4 0.9 × 0.8 0.8 × 0.8 0.7 × 0.7 0.6 × 0.6 0.5 × 0.6 3 1.7 × 1.1 1.7 × 1.1 1.7 × 1.0 1.7 × 1.1 1.3 × 0.7 0.7 × 0.6 0.5 × 0.5 0.4 × 0.4 0.3 × 0.3 0.2 × 0.3 four 1.6 × 1.1 1.6 × 1.1 1.6 × 1.0 1.6 × 1.1 1.2 × 0.8 0.7 × 0.6 0.5 × 0.5 0.4 × 0.4 0.3 × 0.3 0.2 × 0.3 5 1.8 × 1.1 2.0 × 1.1 1.7 × 1.0 1.8 × 1.1 1.1 × 0.8 0.5 × 0.5 0.3 × 0.3 0.2 × 0.2 0.1 × 0.1 No signs of tissue necrosis 6 1.3 × 1.3 1.5 × 1.3 1.3 × 1.3 1.3 × 1.3 1.2 × 0.5 0.6 × 0.4 0.2 × 0.2 0.1 × 0.2 0.1 × 0.1 7 1.8 × 1.1 1.8 × 1.1 1.6 × 1.1 1.8 × 1.1 1.5 × 0.6 0.7 × 0.5 0.6 × 0.3 0.5 × 0.2 0.4 × 0.1 0.3 × 0.1 8 1.6 × 1.0 1.8 × 1.0 1.6 × 1.0 1.6 × 1.0 1.3 × 0.7 0.7 × 0.6 0.6 × 0.5 0.5 × 0.4 0.5 × 0.3 0.4 × 0.2 Experience: Model of a chemical burn (0.3 ml of a 40% solution of formalin intradermally in the back) + treatment with hydrochloric mud extract (according to the scheme indicated in the text) After each measurement, a dirt extract is applied to the defect zone, previously rubbed one 1.1 × 1.2 1.0 × 0.8 0.8 × 0.6 No signs of tissue necrosis No signs of tissue necrosis No signs of tissue necrosis 2 1.1 × 1.6 0.9 × 0.8 0.7 × 0.7 3 1.7 × 1.0 0.7 × 0.6 0.4 × 0.4 four 1.6 × 1.0 0.7 × 0.6 0.4 × 0.4 5 1.4 × 1.0 0.5 × 0.5 0.2 × 0.2 6 1.3 × 1.3 0.6 × 0.4 0.1 × 0.2 7 1.6 × 1.1 0.7 × 0.5 0.5 × 0.2 0.3 × 0.1 8 1.6 × 1.0 0.7 × 0.6 0.5 × 0.4 0.3 × 0.2 0.1 × 0.1 9 1.5 × 0.7 0.5 × 0.5 0.2 × 0.2 0.2 × 0.1 0.2 × 0.1 10 1.4 × 1.2 0.6 × 0.4 0.1 × 0.2 0.1 × 0.2 0.1 × 0.2 Experience:

Model of a chemical burn (0.3 ml of a 40% solution of formalin intradermally in the back) + treatment with an aqueous mud extract (according to the scheme indicated in the text) After each measurement, a dirt extract is applied to the defect zone, previously rubbed one 1.2 × 1.2 1.3 × 1.2 1.2 × 1.2 1.0 × 1.0 1.0 × 0.8 0.5 × 0.7 No signs of tissue necrosis No signs of tissue necrosis No signs of tissue necrosis 2 1.1 × 1.6 1.1 × 1.6 1.1 × 1.6 0.9 × 1.4 0.9 × 0.8 0.4 × 0.6 3 1.7 × 1.1 1.7 × 1.0 1.7 × 1.1 1.3 × 0.7 0.7 × 0.6 No signs of tissue necrosis four 1.5 × 1.1 1.6 × 1.0 1.6 × 1.1 1.2 × 0.8 0.7 × 0.6 5 1.6 × 1.1 1.7 × 1.0 1.8 × 1.1 1.1 × 0.8 0.5 × 0.5 6 1.3 × 1.3 1.3 × 1.3 1.3 × 1.3 1.2 × 0.5 0.6 × 0.4 7 1.5 × 1.1 1.6 × 1.1 1.8 × 1.1 1.5 × 0.6 0.7 × 0.5 0.6 × 0.3 0.2 × 0.2 8 1.6 × 1.0 1.6 × 1.0 1.6 × 1.0 1.3 × 0.7 0.7 × 0.6 0.6 × 0.5 0.7 × 0.7 9 1.2 × 1.2 1.3 × 1.2 1.2 × 1.2 1.0 × 1.0 1.0 × 0.8 0.9 × 0.7 0.5 × 0.2 0.3 × 0.1 10 1.1 × 1.6 1.1 × 1.6 1.1 × 1.6 0.9 × 1.4 0.9 × 0.8 0.8 × 0.8 0.5 × 0.4 0.4 × 0.2

Claims (1)

A method of obtaining a mud extract with a wound healing effect, including extraction of mud, sedimentation, filtration and sterilization, characterized in that the native peloid raw material is mud source - mud "Lake" Healing "with a natural moisture content of 38%, 1 kg of which is subjected to extraction at three times pouring in 3-4 volumes of an aqueous solution of hydrochloric acid (1: 1) at a temperature of 25-30 ° C with vigorous stirring until a homogeneous suspension is obtained, followed by 12 hours of settling at room temperature, filtration micelles, distillation of water on a rotary evaporator at 50-60 ° C and a residual pressure of 10-15 mm Hg and sterilization for 2-3 hours at 125-130 ° C.